Brief Report: Autophagy during beef aging

The conversion of muscle into meat is a complex process of major concern for meat scientists due to its influence on the final meat quality. The aim of this study was to investigate the occurrence of autophagic processes in the conversion of muscle into meat. Our findings demonstrated, for the first time, the occurrence of autophagic processes in the muscle tissue at early postmortem period (2 h to 24 h) in both beef breeds studied (Asturiana de los Valles and Asturiana de la Montaña) showing significant time-scale differences between breeds, which could indicate a role of this process in meat maturation. These breeds have different physiological features: while Asturiana de los Valles is a meat-specialized breed showing high growth rate, an elevated proportion of white fibers in the muscle and low intramuscular fat level, Asturiana de la Montaña is a small- to medium-sized rustic breed adapted to less-favored areas, showing more red fibers in the muscle and a high intramuscular fat content.     

Genetic relationships between seven Spanish native breeds of cattle

Summary. Ten genetic markers were studied in seven Spanish native cattle breeds, using a total of 725 animals. Of the ten, two were found to be monomorphic in all seven breeds. The genetic relationships of the seven breeds are estimated by three different genetic-statistical methods (genetic distances, main coordinate analysis and cluster analysis), which indicate three clearly distinct groups of populations: one where the Cárdena Andaluza and Alistana Sanabresa are very closely related, one comprising Sayaguesa, Morucha, Asturiana de los Valles and Asturiana de la Montaña cattle, and a third, genetically distant from the other two, comprising only Blanca Cacereña. The dendrogram drawn from the genetic distances matrix would seem to imply that the seven breeds are descended from different ancestors.     

Pedigree analysis of eight Spanish beef cattle breeds

The genetic structure of eight Spanish autochthonous populations (breeds) of beef cattle were studied from pedigree records. The populations studied were: Alistana and Sayaguesa (minority breeds), Avileña – Negra Ibérica and Morucha ("dehesa" breeds, with a scarce incidence of artificial insemination), and mountain breeds, including Asturiana de los Valles, Asturiana de la Montaña and Pirenaica, with extensive use of AI. The Bruna dels Pirineus breed possesses characteristics which make its classification into one of the former groups difficult. There was a large variation between breeds both in the census and the number of herds. Generation intervals ranged from 3.7 to 5.5 years, tending to be longer as the population size was larger. The effective numbers of herds suggest that a small number of herds behaves as a selection nucleus for the rest of the breed. The complete generation equivalent has also been greatly variable, although in general scarce, with the exception of the Pirenaica breed, with a mean of 3.8. Inbreeding effective population sizes were actually small (21 to 127), especially in the mountain-type breeds. However, the average relatedness computed for these breeds suggests that a slight exchange of animals between herds will lead to a much more favourable evolution of inbreeding. The effective number of founders and ancestors were also variable among breeds, although in general the breeds behaved as if they were founded by a small number of animals (25 to 163).     

Effect of age at weaning on the physiological stress response and temperament of two beef cattle breeds

The study was conducted to evaluate the effect of age at weaning and breed on the stress response of calves to weaning and their temperament. At calving, 14 Parda de Montaña calves and 14 Pirenaica calves were randomly assigned to either early weaning (at 90 days) or traditional weaning (at 150 days) treatment. During nursing, calves were allowed to suckle their dams twice a day for 30 min. After weaning, calves were placed in an adjacent barn without access to their dams, where they remained for 7 days. On day 8 after weaning, they were transported to a feedlot where they received an intensive diet. Blood samples were taken 168 h before weaning (baseline) and 6, 24, 48 and 168 h after weaning for cortisol, fibrinogen and haematology analyses, and temperament was measured 90 and 180 days after weaning with the flight speed test. Cortisol concentration increased after weaning, irrespective of age at weaning. Early-weaned calves had a lower fibrinogen baseline and a greater increase in fibrinogen concentrations 48 h after weaning than traditionally weaned calves. Moreover, fibrinogen concentration returned to baseline values 168 h after weaning in traditionally weaned calves, whereas it remained high in early-weaned calves. Concerning breed effects, Pirenaica calves had higher cortisol concentration and fibrinogen increments after weaning than Parda de Montaña calves. Slight alterations occurred after weaning in haematology, but all parameters returned to baseline values 168 h after weaning, with no significant effects of age at weaning or breed. Despite the absence of clinical signs, early-weaned calves of both breeds suffered marginal anaemia, according to haemoglobin values. Regardless of age at weaning, Pirenaica calves had greater reactivity to human presence than Parda de Montaña calves, according to their higher flight speed values measured. Finally, early-weaned calves were lighter than traditionally weaned calves at weaning, but had similar weight gains in the feedlot. Consequently, they needed an additional 40 days to reach the target weight, irrespective of breed. Therefore, age at weaning had no major effects on the stress response to weaning or temperament, but early weaning increased the length of the feedlot period. On the other hand, Pirenaica calves were more reactive than Parda de Montaña calves to the stress of weaning and human presence.     

Effect of weight at slaughter and breed on beef intramuscular lipid classes and fatty acid profile

The effect of slaughter weight and breed on the composition of intramuscular fat (IMF) of seven Spanish cattle breeds was studied. Lipid classes and fatty acids were evaluated in young bulls of seven local Spanish cattle breeds, Asturiana, Avileña, Morucha, Parda Alpina, Pirenaica, Retinta and Rubia Gallega, slaughtered at a live weight of 320 kg (veal type) and 550 kg (yearling bull type). Higher slaughter weight and early maturating breeds produced higher IMF content in the longissimus muscle (P < 0.001), which was linked to a reduction in phospholipids, monoglycerides, diglycerides, cholesterol and free-fatty acids content, but an increase in triacylglycerols. Besides, heavier animals displayed a higher percentage of saturated and monounsaturated fatty acids (MUFA), but a lower polyunsaturated fatty acids (PUFA) percentage (P < 0.001). The increase in triglycerides (TG), saturated fatty acids (SFA) and MUFA illustrates the increasing importance of lipid storage as fattening proceeds and the acquisition of ruminal functionality. The feeding system based on cereals had a strong influence on the fatty acid profile, giving the studied beef a relatively high PUFA/SFA and n-6/n-3 ratios.     

Assessing priorities for conservation in Tuscan cattle breeds using microsatellites

Preservation of rare genetic stocks requires assessment of within-population genetic diversity and between-population differentiation to make inferences on their degree of uniqueness. A total of 194 Tuscan cattle (44 Calvana, 35 Chianina, 25 Garfagnina, 31 Maremmana, 31 Mucca Pisana and 28 Pontremolese) individuals were genotyped for 34 microsatellite markers. Moreover, 56 samples belonging to Argentinean Creole and Asturiana de la Montaña cattle breeds were used as an outgroup. Genetic diversity was quantified in terms of molecular coancestry and allelic richness. STRUCTURE analyses showed that the Tuscan breeds have well-differentiated genetic backgrounds, except for the Calvana and Chianina breeds, which share the same genetic ancestry. The between-breed Nei's minimum distance (D_m) matrices showed that the pair Calvana–Chianina was less differentiated (0.049 ± 0.006). The endangered Tuscan breeds (Calvana, Garfagnina, Mucca Pisana and Pontremolese) made null or negative contributions to diversity, except for the Mucca Pisana contribution to allelic richness (C_T = 1.8%). The Calvana breed made null or negative within-breed contributions (f¯_W = 0.0%; C_W = −0.4%). The Garfagnina and Pontremolese breeds made positive contributions to between-breed diversity but negative and high within-breed contributions, thus suggesting population bottleneck with allelic losses and increase of homozygosity in the population. Exclusion of the four endangered Tuscan cattle breeds did not result in losses in genetic diversity (f¯_T = −0.7%; C_T = −1.2%), whereas exclusion of the non-endangered breeds (Chianina and Maremmana) did (f¯_T = 2.1%; C_T = 3.9%); the simple exclusion of the Calvana breed from the former group led to losses in genetic diversity (f¯_T = 0.47%; C_T = 2.34%), indicating a diverse significance for this breed. We showed how quantifying both within-population diversity and between-population differentiation in terms of allelic frequencies and allelic richness provides different and complementary information on the genetic backgrounds assessed and may help to implement priorities and strategies for conservation in livestock.     

Characterization of the Myofibrillar Proteome as a Way to Better Understand Differences in Bovine Meats Having Different Ultimate pH Values

Influence of ultimate pH (pHu) on the occurrence of defective meats known as dark, firm, and dry (DFD) meats has been studied through a proteomic approach at early post‐mortem times. The myofibrillar sub‐proteome of longissimus thoracis et lumborum muscle from twelve loin samples from Asturiana de los Valles x Friesian yearling bulls, previously classified into two groups of six samples according to their pH values (normal, pHu < 6.0 and high, pHu ≥ 6.0), is analyzed at 24 h post‐mortem. Fractionation/enrichment of muscle samples is carried out by combining OFFGEL fractionation in the pH range 4–7 followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE) of the retrieved liquid fractions. Four protein bands satisfactorily discriminate between meat samples with normal and high pHu. These bands are quantified by image analysis, and further identified by liquid chromatography‐mass spectrometry as desmin, pyruvate kinase, myosin light chain, and myosin heavy chain‐1 and ‐2. Coupling OFFGEL and SDS‐PAGE separation with MS provides detailed and reproducible myofibrillar protein profiles enabling comparison among the sample groups assayed. This makes feasible to identify biomarkers capable to better understand pre‐slaughter stress condition susceptible to give DFD meats with high pHu values.     

Comparison of microRNAs in adipose and muscle tissue from seven indigenous Chinese breeds and Yorkshire pigs

Elucidation of the pig microRNAome is essential for interpreting functional elements of the genome and understanding the genetic architecture of complex traits. Here, we extracted small RNAs from skeletal muscle and adipose tissue, and we compared their expression levels between one Western breed (Yorkshire) and seven indigenous Chinese breeds. We detected the expression of 172 known porcine microRNAs (miRNAs) and 181 novel miRNAs. Differential expression analysis found 92 and 12 differentially expressed miRNAs in adipose and muscle tissue respectively. We found that different Chinese breeds shared common directional miRNA expression changes compared to Yorkshire pigs. Some miRNAs differentially expressed across multiple Chinese breeds, including ssc‐miR‐129‐5p, ssc‐miR‐30 and ssc‐miR‐150, are involved in adipose tissue function. Functional enrichment analysis revealed that the target genes of the differentially expressed miRNAs are associated mainly with signaling pathways rather than metabolic and biosynthetic processes. The miRNA–target gene and miRNA–phenotypic traits networks identified many hub miRNAs that regulate a large number of target genes or phenotypic traits. Specifically, we found that intramuscular fat content is regulated by the greatest number of miRNAs in muscle tissue. This study provides valuable new candidate miRNAs that will aid in the improvement of meat quality and production.     

Skeletal muscle transcriptional profiles in two Italian beef breeds, <Emphasis Type="Italic">Chianina</Emphasis> and <Emphasis Type="Italic">Maremmana</Emphasis>, reveal breed specific variation

Chianina and Maremmana breeds play an important role in the Italian cattle meat market. The Chianina breed is an ancient breed principally raised for draught. Now this breed is the worldwide recognized producer of top quality beef, tasteful and tender, specifically the famous “Florentine steak”. The Maremmana characterized by a massive skeletal structure, is a rustic cattle breed selected for adaptability to the marshy land of the Maremma region. We used a high throughput mRNA sequencing to analyze gene expression in muscle tissues of two Italian cattle breeds, Maremmana (MM) and Chianina (CN) with different selection history. We aim to examine the specific genetic contribution of each breed to meat production and quality, comparing the skeletal muscle tissue from Maremmana and Chianina. Most of the differentially expressed genes were grouped in the Glycolysis/Gluconeogenesis pathways. The rate and the extent of post-mortem energy metabolism have a critical effect on the conversion of muscle to meat. Furthermore, we aim at discovering the differences in nucleotide variation between the two breeds which might be attributable to the different history of selection/divergence. In this work we could emphasize the involvement of pathways of post-mortem energy metabolism. Moreover, we detected a collection of coding SNPs which could offer new genomic resources to improve phenotypic selection in livestock breeding program.     

Transcriptional alteration of p53 related processes as a key factor for skeletal muscle characteristics in <Emphasis Type="Italic">Sus scrofa</Emphasis>

The pig could be a useful model to characterize molecular aspects determining several delicate phenotypes because they have been bred for those characteristics. The Korean native pig (KNP) is a regional breed in Korea that was characterized by relatively high intramuscular fat content and reddish meat color compared to other western breeds such as Yorkshire (YS). YS grew faster and contained more lean muscle than KNP. We compared the KNP to Yorksire to find molecular clues determining muscle characteristics. The comparison of skeletal gene expression profiles between these two breeds showed molecular differences in muscle. We found 82 differentially expressed genes (DEGs) defined by fold change (more than 1.5 fold difference) and statistical significance (within 5% of false discovery rate). Functional analyses of these DEGs indicated up-regulation of most genes involved in cell cycle arrest, down-regulation of most genes involved in cellular differentiation and its inhibition, down-regulation of most genes encoding component of muscular-structural system, and up-regulation of most genes involved in diverse metabolism in KNP. Especially, DEGs in above-mentioned categories included a large number of genes encoding proteins directly or indirectly involved in p53 pathway. Our results indicated a possible role of p53 to determine muscle characteristics between these two breeds.     

The rearing system modulates biochemical and histological differences in loin and ham muscles between Basque and Large White pigs

Conventional pork production, based on highly selected breeds for growth efficiency and carcass leanness, is generally considered to decrease pork quality. In contrast, non-selected breeds produced in extensive systems are associated with high pork quality, which is generally attributed to higher intramuscular fat (IMF) content and less glycolytic muscle metabolism. The present study aimed to determine biochemical, histological and quality traits of loin and ham muscles of pigs from selected Large White (LW) and local French, non-selected Basque (B) breeds. Pigs were reared in a conventional indoor (C, slatted floor), alternative (A, indoor bedding and outdoor area) or extensive system (E, free range, B pigs only). A total of 100 castrated males were produced in 2 replicates, each containing 5 groups of 10 pigs based on breed and system: LWC, LWA, BC, BA and BE. The glycolytic longissimus muscle (LM) and semimembranosus muscle (SM), and the deep red (RSTM) and superficial white (WSTM) portions of semitendinosus muscle (STM) were studied at 145 kg BW. Overall, breed induced stronger effects on muscle traits than the rearing system, among which the E system induced greater changes. The lower muscle growth of B pigs was associated with fewer muscle fibers and a smaller cross-sectional area (CSA) of glycolytic fibers (P < 0.01). The SM was less glycolytic and more oxidative in B than in LW pigs (P < 0.001). The WSTM followed a similar trend, with a larger relative area of type I fibers in B pigs. In contrast, the LM and RSTM were more oxidative in LW pigs. B pigs had higher IMF content and ultimate pH in all muscles, along with lower glycolytic potential, less light and redder meat in the LM and SM (P < 0.001). Compared to the C system, the A system induced only a shift towards a more oxidative metabolism in the LM and a smaller fiber CSA in the RSTM of LW pigs (P < 0.05), without influencing pork quality traits. Compared to BC pigs, BE pigs had a more oxidative and less glycolytic muscle metabolism, along with higher ultimate pH, lower lightness and redder meat (P < 0.01), but similar IMF content. Overall, results indicate that influences of breed and rearing system on muscle properties depend on muscle type, and that IMF content and fiber-type composition are unrelated traits that can be modified independently by genetic or rearing factors.     

Quality traits of suckling-lamb meat covered by the protected geographical indication “Lechazo de Castilla y León” European quality label

The quality of suckling-lamb meat (derived from Churra, Castellana and Ojalada breeds), covered by the protected geographical indication (PGI) “Lechazo de Castilla y León”, and the effects of carcass weight (CW), breed and sex on quality were studied. The edible portion (EP) of 81 carcasses was analysed for proximate composition, fatty acid, cholesterol, amino acid and mineral contents. The longissimus thoracis and lumborum muscle (30 samples) were analysed for proximate composition, pH, myoglobin content, water holding capacity, Warner–Bratzler shear force, and total and soluble collagen content. This study contributes to characterization of suckling-lamb meat quality recognised in the European Union with a PGI label and provides new data on the composition of the EP of the carcasses. CW and breed had a significant effect on several quality traits, most related to fatness.     

Exploring alternative models for sex-linked quantitative trait loci in outbred populations: application to an iberian x landrace pig intercross

We present a very flexible method that allows us to analyze X-linked quantitative trait loci (QTL) in crosses between outbred lines. The dosage compensation phenomenon is modeled explicitly in an identity-by-descent approach. A variety of models can be fitted, ranging from considering alternative fixed alleles within the founder breeds to a model where the only genetic variation is within breeds, as well as mixed models. Different genetic variances within each founder breed can be estimated. We illustrate the method with data from an F(2) cross between Iberian x Landrace pigs for intramuscular fat content and meat color component a*. The Iberian allele exhibited a strong overdominant effect for intramuscular fat in females. There was also limited evidence of one or more regions affecting color component a*. The analysis suggested that the QTL alleles were fixed in the Iberian founders, whereas there was some evidence of segregation in Landrace for the QTL affecting a* color component.     

Effect of low- and high-forage diets on meat quality and fatty acid composition of Alentejana and Barrosã beef breeds

This study investigated the effects of genotype and diet on meat fat composition and palatability obtained from Alentejana (AL) and Barrosã (BA) breeds. Herein, 20 males from each breed allocated at 11 months of age were fed ad libitum a low-forage diet or a high-forage diet and slaughtered at 18 months of age. Trained sensory panel analysis found that the longissimus lumborum (Ll) muscle from BA had higher tenderness, juiciness and overall acceptability scores than the AL breed. The highest scores for those attributes were observed in the BA breed fed the high-forage diet. Regarding the semitendinosus (St) muscle, breed was a source of variation of tenderness scores. In contrast to the Ll muscle, the highest tenderness scores for the St muscle were observed in the AL breed. The intramuscular fat (IMF) content was positively correlated with tenderness, juiciness and overall acceptability in Ll muscle and negatively correlated with flavour in the St muscle. The levels of 14:0 and 16:0, 16:1c9, 18:1c9 and 18:1c11 were positively correlated to juiciness, tenderness and overall acceptability in the Ll muscle. These correlations were not observed in the St muscle, which may be related to its low IMF content. Nonetheless, negative correlations were observed for the St muscle between flavour and 14:0, 16:0 and 18:0 FA contents.The IMF varied widely in the Ll but not in the St muscle. The latter had higher levels of 16:1c9 and trans fatty acids (∑TFA) in the BA than in the AL breed. Regarding the Ll muscle, the BA had higher amounts of 14:0, 16:0, 16:1c9, 18:0, 18:1c9, 18:1c11, saturated fatty acids (∑SFA), cis monounsaturated fatty acids (∑cis MUFA), ∑TFA and n-3 polyunsaturated fatty acids (∑n-3 PUFA) than the AL breed. The diet exerted an influence on the IMF content and on the levels of 14:0, 16:0, 16:1c9, 18:0, 18:1c9, 18:1c11, ∑SFA, ∑cis MUFA and ∑TFA in both Ll and St muscles. Moreover, the levels of ∑n-3 PUFA in the Ll muscle and 18:2n-6, 20:4n-6, ∑n-6 PUFA and ∑PUFA in the St muscle were influenced by diet. The results obtained in this study, with two Portuguese breeds, confirm that genetic background plays a major role in the determination of meat eating quality.     

Effect of myosin heavy chain composition of muscles on meat quality in Laiwu pigs and Duroc

In order to explain the mechanism of high meat quality in Laiwu pigs and investigate the relation between myosin heavy chains (MyHC) composition and meat quality, meat quality analysis was conducted and mRNA expression of MyHC I, IIa, IIx, IIb was quantified by real-time fluorescence PCR in longissimus muscle (LM) and semimembranous muscle of Laiwu pigs and Duroc. The result indicated that, compared with Duroc, mRNA expression of MyHC IIa, IIx in LM and semimembranous muscle of Laiwu pigs was significantly increased, mRNA expression of MyHC IIb was dramatically decreased. However, the expression of MyHC I was not significantly affected by breeds. The correlation between mRNA expression of MyHC I, IIa, IIx in LM and meat color, pH value, marbling, intramuscular fat content was positive, but shear value of LM was negative. The relation between MyHC IIb mRNA expression and marbling, intramuscular fat content was dramatically negative, whereas shear value was strikingly positive, as well as fiber diameter, but without reaching statistical significance. Therefore, the composition of MyHC I, IIa, IIx, IIb affected meat quality, furthermore, expression of MyHC I, IIa, IIx, IIb mRNA prominently influenced meat characteristics, especially edible quality of muscle, suggesting that mRNA expression level of MyHC I, IIa, IIx, IIb can exactly and impersonally estimate meat quality.